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Method Article
Jason C. Klima
University of Washington
Corresponding Author
ORCiD: https://orcid.org/0000-0003-0387-7847
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Bacterial expression and purification of de novo designed mini-fluorescence-activating proteins (mFAPs) are accomplished by genetically fusing mFAP variants to an N-terminal 6xHis tag and optionally to a Tobacco Etch Virus (TEV) protease epitope. The protocol can take less than 24 hours to complete, and allows for downstream in vitro characterization of photophysical properties of mFAPs by equilibration with exogenous fluorogenic compounds.
Keywords
bacterial expression, protein purification
This is a list of supplementary files associated with this preprint. Click to download.
- TEVprotease.fasta
Supplementary File 1. Tobacco Etch Virus (TEV) protease amino acid sequence used in this protocol in FASTA format.
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Associated Publications
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Jason C. Klima
University of Washington
Corresponding Author
ORCiD: https://orcid.org/0000-0003-0387-7847
Version History
CURRENT STATUS: Posted
Version 1
Posted 08 Feb, 2021
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Bacterial expression and purification of de novo designed mini-fluorescence-activating proteins (mFAPs) are accomplished by genetically fusing mFAP variants to an N-terminal 6xHis tag and optionally to a Tobacco Etch Virus (TEV) protease epitope. The protocol can take less than 24 hours to complete, and allows for downstream in vitro characterization of photophysical properties of mFAPs by equilibration with exogenous fluorogenic compounds.
This is a list of supplementary files associated with this preprint. Click to download.
- TEVprotease.fasta
Supplementary File 1. Tobacco Etch Virus (TEV) protease amino acid sequence used in this protocol in FASTA format.
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